Pepsin Immobilized on High-Strength Hybrid Particles for Continuous Flow Online Digestion at 10 000 psi
| العنوان: | Pepsin Immobilized on High-Strength Hybrid Particles for Continuous Flow Online Digestion at 10 000 psi |
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| المؤلفون: | Joomi Ahn, Ying Qing Yu, Kevin D. Wyndham, John R. Engen, Moon Chul Jung |
| المصدر: | Analytical Chemistry. 84:7256-7262 |
| بيانات النشر: | American Chemical Society (ACS), 2012. |
| سنة النشر: | 2012 |
| مصطلحات موضوعية: | Silicon dioxide, Proteolysis, Molecular Sequence Data, Mass spectrometry, Article, Mass Spectrometry, Analytical Chemistry, chemistry.chemical_compound, Digestion (alchemy), Pepsin, Pressure, medicine, Animals, Amino Acid Sequence, Particle Size, Chromatography, medicine.diagnostic_test, biology, Deuterium Exchange Measurement, Enzymes, Immobilized, Silicon Dioxide, Pepsin A, Peptide Fragments, chemistry, biology.protein, Isobaric process, Particle size |
| الوصف: | Pepsin was immobilized on ethyl-bridged hybrid (BEH) particles, and digestion performance was evaluated in a completely online format, with the specific intent of using the particles for hydrogen-deuterium exchange mass spectrometry (HDX MS) experiments. Because the BEH particles are mechanically strong, they could withstand prolonged, continuous high-pressure at 10,000 psi. Online digestion was performed under isobaric conditions with continuous solvent flow, in contrast to other approaches where the pressure or flow is cycled. As expected, digestion efficiency at 10,000 psi was increased and reproducibly produced more peptic peptides versus digestion at 1000 psi. Prototype columns made with the BEH pepsin particles exhibited robust performance, and deuterium back-exchange was similar to that of other immobilized pepsin particles. These particles can be easily incorporated in existing HDX MS workflows to provide more peptide coverage in experiments where fast, efficient, and reproducible online pepsin digestion is desired. |
| تدمد: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac301749h |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c902dcaaf0f541a4927db47bb627c41b https://doi.org/10.1021/ac301749h |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....c902dcaaf0f541a4927db47bb627c41b |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15206882 00032700 |
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| DOI: | 10.1021/ac301749h |